Figure 3. The expression of Notch pathway genes are dependent of Apc status in HT29 cells and murine intestinal adenomas.

Wt-Apc was induced in HT29-APC cells through the addition of 100 µM ZnCl₂ to the growth medium. Semi-quantitative RT-PCR was carried out on cDNA reversely transcribed from 200 ng total RNA from each time point (0–24 h post wt-Apc induction). Bars describe the relative expression of Notch-2 in HT29-APC cells (*) versus HT29-β-galactosidase cells (**) normalized against Gapdh expression. Cyclin D1 was used as a positive control for Wnt inactivation. (B) The protein expression of Apc (∼310 kDa), Notch-2 (∼265 kDa), wt and loading control, Gapdh, was determined with Western blot following 0–24 h of zinc stimulation. Adjustments in whole image contrast levels were performed in Adobe Photoshop CS4. (C) Relative mRNA expression of murine Notch-1, Notch-2 and Hes-1 in tumors and corresponding non-tumor normal intestinal mucosa of Apc^Min/+ mice. mRNA expression was related to the endogenous control gene GAPDH. White columns (n = 16), black columns (n = 22). Bars are presented as median expression values. Error bars describe interquartile range.